EP2830763B1 - Integrated disposable chip cartridge system for mobile multiparameter analyses of chemical and/or biological substances - Google Patents

Description

The invention relates to a disposable measuring cartridge module (in the following also briefly: measuring cartridge module, measuring cartridge or cartridge) for carrying out preferably mobile multiparameter analyzes of chemical and / or biological substances, in particular for the detection and / or determination of the concentration of target substances in liquid samples. Furthermore, the invention relates to a disposable chip cartridge system (hereinafter referred to as "system") comprising an exchangeable measuring cartridge cartridge module and a method for performing such analyzes in such a system.

Systems of this type are used in the field of mobile reproducible chemical and biochemical analyzes, in particular in the field of point-of-care diagnostics. The aim is to forego conventional stationary laboratory analysis with unavoidable waiting times and instead allow for a method of obtaining the desired on-site analysis results, e.g. at the place of patient treatment, immediate and prompt (virtually immediate) receives.

• WO 2006/125767 A1 offenbart ein Analysegerät zur integrierten und automatisierten DNA- oder Protein-Analyse, das in ein Steuergerät und eine in dieses einsetzbare Cartridge in Form und Größe einer Scheckkarte aufgeteilt ist. Die Cartridge enthält Kanalabschnitte mit diversen Reagenzien in trockener und langzeitstabiler Form, einen Probenport zum Einpipettieren einer Probe, eine PCR-Kammer zur Durchführung einer PCR (polymerase chain reaction) und eine Detektionskammer mit einem DNA-Sensor. Das für den Analysevorgang erforderliche Wasser wird im Steuergerät aufbewahrt und mittels fluidischer Komponenten des Steuergeräts, unter anderem einer Pumpe und diverser Füllstandssensoren, zu den Wasserports der in das Steuergerät eingesetzten Cartridge befördert. Die Steuerung der Wasserbeförderung sowie das Auslesen der Messdaten erfolgt mittels einer ebenfalls im Steuergerät untergebrachten Elektronik.

The following systems are known from the prior art, which are based on interchangeable measuring cartridges for receiving and analyzing a sample:

• WO 2006/125767 A1 discloses an analysis device for integrated and automated DNA or protein analysis, which is divided into a control unit and a usable therein cartridge in the form and size of a check card. The cartridge contains channel sections with various reagents in dry and long-term stable form, a sample port for pipetting a sample, a PCR chamber for performing a PCR (polymerase chain reaction) and a detection chamber with a DNA sensor. The water required for the analysis process is stored in the control unit and conveyed by means of fluidic components of the control unit, including a pump and various level sensors, to the water ports of the cartridge inserted in the control unit. The control of the transport of water as well as the readout of the measured data are carried out by means of an electronic unit also housed in the control unit.

US 2007/0166192 A1 shows a sample preparation module for receiving and processing specific biological samples for analysis in a standardized and miniaturized "lab-on-a-chip" cartridge. This module is designed for a specific sample type and has a cavity for complex or large-scale preparation steps that can not be performed inside the cartridge. These are, for example, extraction or multiplication of cells of a target substance, mechanical homogenization of solid tissue samples, etc. The prepared sample passes out of the module via a suitably dimensioned and sealed opening directly into the sample entrance of the cartridge. The Sampling and dispensing is performed in the module either manually and by using gravity or by means of a displaceable piston in the cavity.

US 5,096,669 discloses a system for performing electrochemical measurements on blood and other fluids with a disposable measuring unit that can be plugged into a mobile reader. The measuring unit contains sensors, a bag with calibration fluid including a spike, an air bubble reservoir and a channel system with sample inlet. The sample passes through the capillary into the channel system. The puncture of the calibration bag for calibration measurement and the transport of the sample to the sensor by the compression of the air bubble are triggered or controlled by the read-out device.

US 5,638,828 shows a sampling device for collecting liquid samples and their introduction into a disposable measuring device for real-time analysis. Within the sampling device, the sample is first collected in a reservoir into which a precisely dimensioned capillary opens, which can be filled with the sample by capillary action. After manually connecting the sampler with the filled capillary to the meter, it is connected at both ends to the inside of the meter. The movement of the sample liquid inside the meter takes place by deliberately compressing an air bubble provided therein, which can move the sample along the capillary and furthermore via the sensor.

US 2003/0170881 A1 discloses an apparatus for performing various analyzes on liquid samples. The adaptability to different analytical procedures is ensured by "valve means" provided within a disposable cartridge, such as constrictions, which influence the flow of the sample and a second calibrating and rinsing liquid including air bubbles provided in a bag in the cartridge. The cartridge is inserted into a read-out device, whereby pressure is exerted on certain paddle-shaped regions in the cartridge cover for transporting the fluids contained in the cartridge. Calibration measurements are taken before or after sample measurement, with channels and sensors in between being cleaned with the second fluid. Reagents are provided in dry form in the channels of the cartridge.

US 5,405,510 relates to a portable measuring unit for the measurement of multiple, sequential fluid samples. The device has a disposable cartridge having access to an inlet to a hydrated sensor flow cell and, in fluid contact but spatially separated, a reference electrode. In addition to the necessary electrical equipment, the cartridge contains a stretchable waste container. Between the waste container and the flow cell is a check valve. After calibration of the cell, a sample may be passed through the inlet with an unknown amount of a known analyte.

In DE 102 45 845 A1 discloses a measuring chip with a fluid overflowable reaction surface (analysis cell), which allows a luminescence reaction and their measurement. The measuring chip itself has, in addition to the analysis cell, spaces which serve to store sample liquid and reagents, which are sealed with a seal or a septum and can be punctured with a syringe. However, it is not a self-contained unit, but cooperates with a base plate, are arranged in valves controllable fluid channels that can connect the pantries and the analysis cell with each other.

In addition, please refer to the following documents: DE 10 1009 007230 A1 discloses a measuring cell having a flexible sidewall component which simultaneously seals to a sensor chip and to a cover. Out DE 41 39 121 C1 the integration of the flow cell into a measuring insert is known, which is interchangeable used in a comprehensive the remaining components of the device base unit. US 2011/0301047 A1 proposes to provide a valve between the inlet and the measuring chamber of a detection device. EP 1 245 285 A2 discloses a syringe-loadable sample inlet. In WO 2009/062940 A1 a sensor cassette is described from two firmly interconnected, but separately manufactured modules and with a continuous measuring channel for receiving fluidic media; Each of the modules is electrically connected via contact with an analyzer. The flow channels of a device for exchangeable recording of measuring cartridges or measuring cells according to WO 98/05958 A1 have repeatedly pierceable septa; In addition, the device has a built-in a valve block sample receptacle, which is accessible from the outside for syringes or capillaries.

The Fraunhofer Institute for Silicon Technology (ISIT), together with POCDIA GmbH, has presented a new point-of-care diagnostic system (https://www.isit.fraunhofer.de/ en / News / Point-of-Care_Diagnostik_System.html; found on 23.07.2013)

• chargenspezifische Kalibrierung durch exemplarische Tests während/nach der Produktion;
• manuelle Arbeitsschritte bei der Nutzung, insbesondere bei der Probenvorbereitung;
• spezielle Proben-Aufreinigung (insbesondere für optische Detektionsmethoden);

und/oder es handelt sich bei ihnen um Vorrichtungen, die durch eine oder mehrere der folgenden Komponenten gekennzeichnet sind:

• Analysegeräte mit darin enthaltenen Fluidikkomponenten (Pumpen, Ventilen) zur Prozessierung von Flüssigkeiten im Gerät (d.h. Flüssigkeitskontakt von Gerätekomponenten) oder alternativ z.B. auch manuelle Zuführung von Flüssigkeiten;
• Verbrauchsmittel-Set mit vielen verschiedenen Komponenten und zum Teil komplizierter Handhabung (Pipetten, Reagenzien, Kartuschen, Chips...).

The known devices and analysis methods each require one or more of the following measures:

• Batch-specific calibration through exemplary tests during / after production;
• manual operations during use, especially during sample preparation;
• special sample purification (especially for optical detection methods);

and / or they are devices characterized by one or more of the following components:

• Analyzers with fluidic components contained therein (pumps, valves) for the processing of liquids in the device (ie fluid contact of device components) or alternatively, for example, also manual supply of liquids;
• Consumables set with many different components and sometimes complicated handling (pipettes, reagents, cartridges, chips ...).

• keine Kalibrierungsmöglichkeit zur Laufzeit der Analyse;
• deutlich begrenzte, auf ein Minimum reduzierte Prozessierungs- und Kontrollmöglichkeiten für die Probe und die Reagenzien innerhalb des Einweg-Messgeräts z.B. mittels einer einzigen Luftblase zur Beförderung sowie Trennung der einzelnen Fluide voneinander;
• Notwendigkeit der Verwendung von Trockenreagenzien und deren Auflösung/Suspension in einem Kanal oder sogar Mikrokanal der Messkartusche während des eigentlichen Analysevorgangs mit damit verbundenen Unsicherheiten bzgl. deren Verteilung, Konzentration und des gesamten Zeitablaufs;
• mangelhafte Güte der Reproduzierbarkeit der Analysen von Test zu Test;
• Erfordernis einer manuellen Probenvorbereitung/-Dosierung vor der Messung z.B. für Blutproben;
• Kontamination von Analysegerätekomponenten, insbesondere der Fluidikkomponenten, durch Flüssigkeitskontakt, was eine aufwendige Wartung und Pflege erfordert;
• eine komplizierte, aufwendige Verbrauchsmittellogistik.

Existing (point-of-care) systems therefore have some of the following disadvantages:

• no calibration option at runtime of the analysis;
• significantly limited, minimized processing and control capabilities for the sample and reagents within the disposable meter, for example by means of a single air bladder for transport and separation of the individual fluids from one another;
• The need to use dry reagents and their dissolution / suspension in a channel or even microchannel of the measuring cartridge during the actual analysis process with associated uncertainties regarding their distribution, concentration and the total time course;
• poor quality of the reproducibility of the analyzes from test to test;
• Requirement of manual sample preparation / dosing prior to measurement, eg for blood samples;
• Contamination of analyzer components, especially the fluidic components, by liquid contact, which requires extensive maintenance and care;
• a complicated, expensive consumables logistics.

Overall, there is a need to increase automation or increase the degree of integration of complete biochemical analysis operations for mobile multiparameter reproducible analyzes, e.g. for the field of point-of-care diagnostics.

The present invention is therefore an object of the invention to provide an apparatus and a method for performing preferably mobile chemical and / or biological sample analyzes, which improved reproducibility of the tests, better automation and increased degree of integration with the lowest possible cost, manufacturing and Allow operating expenses.

This object is achieved by a measuring cartridge module for carrying out preferably mobile multiparameter analyzes of chemical and / or biological substances, comprising a sample inlet, through which a sample substance can pass from the outside into the measuring cartridge module, in addition to one or more reagent inputs, by which reagents from outside, eg from one suitable reagent storage and processing module, the Messchipkartuschenmodul can be supplied, a measuring chip with a provided for sample analysis surface that at least partially represents a bottom or wall of a two-port analysis cell, which is designed such that a liquid comprising the sample substance by a first channel of the analysis cell can be supplied, as well as a channel system in which the sample substance from the sample inlet to the analysis cell and the reagents from the at least one reagent inlet to the analysis cell conveys t can be. The channel system contains in its interior before starting the entire analysis process required for the analysis solid substances (reagents in solid form), and preferably no substances other than air and / or inert gas, which get into the analysis cell can. The measuring cartridge module according to the invention may in particular have the configuration according to claim 1.

The sample substance (in short: sample) can be supplied to the sample inlet in the form of a liquid such as blood or serum, a solid or a wipe sample.

A first reagent inlet externally separated from the sample inlet may be e.g. be used for transport, dilution or liquefaction of the sample substance and / or addition of reactive substances required for analysis. While the sample inlet itself can be tailored to the collection of the sample substance and can usually be closed after a successful sampling, this further inlet into the cartridge makes it possible to flexibly configure the further processing of the sample substance.

The term "reagent" is used in this context generally for any substance that can enter the channel system of the cartridge via a reagent inlet. As reagents, within the scope of the invention, e.g. chemically or biologically active substances, calibration solutions with known concentrations of one or more reference substances, buffer solutions, enzyme solutions, substrate solutions, detector solutions, (e.g., isotonic) salt solutions, distilled water, cleaning solutions, air or inert gas.

In one embodiment, the module of the present invention includes at least two reagent inputs, one of which may be used for the delivery of calibration solution to allow for calibration at run time in a single channel system, as described in more detail below. Of course, more than the aforementioned two reagent inputs may be present in this embodiment.

The term "measurement" and "measuring chip" etc., in the context of the present application, in addition to a possible measurement of one or more parameters such as e.g. the concentration of certain substances e.g. chemical and / or biological nature, such as individual molecules, substances or cells (quantitative measurements), and their qualitative detection.

The measuring chip can have, for example, electrodes, possibly with catcher molecules for a target substance, for carrying out electrochemical analyzes. The electrodes may be formed as inter-digital electrodes; the surface may be occupied by gold or made of gold to facilitate connection of catcher molecules via SH bonds. The measurement can be carried out amperometrically, alternatively voltammetrically or by both methods. It is also an optical measurement for sample analysis possible.

The analysis cell is arranged on the surface of the measuring chip intended for the measurement, for example immediately above a flat measuring chip. In specific embodiments, for example, a bottom of the measuring chip can be used for the measurement. The measuring chip can also be arranged laterally within the cartridge, with a corresponding arrangement of the analysis cell at its intended for measurement inner surface relative to the cartridge.

The analysis cell (in short: cell) is designed to receive a liquid. It contains a first opening through which the liquid can enter the cell, while e.g. Air or spent solution is displaced from the cell through the second opening.

To transport the sample and reagents in the cartridge between the respective exterior inputs (meaning the sample and one or more reagent inputs) to the cell, a channel system is provided. This may include inside the cartridge both interconnected and completely separate subsystems of channels.

The channel system is used e.g. connecting a reagent inlet to the sample inlet within the cartridge to mix the sample with a reagent prior to entering the analysis cell, as previously briefly discussed.

The cartridge of the present invention does not contain any dry reagents in the channel system prior to the addition of reagents from outside via the reagent inputs.

Preferred embodiments of the invention are indicated in the dependent claims and in the following description.

Preferably, the analysis cell is a flow cell, i. a liquid can enter the flow cell through the first opening and leave it partially or completely through the second opening. By applying a suction pressure to the second opening, for example via its connection to a reagent inlet, e.g. the flow or distribution of the liquid to be analyzed over the measuring chip surface are affected.

In addition, a flow cell allows a successive analysis of a reference substance and a sample. Thus, for example, a calibration solution containing a known amount of one or more reference substance (s), passed through one of several reagent inputs in the flow cell and the contained (s) reference substance (s), optionally after connection to corresponding catcher molecules and / or Using other reagents, be determined qualitatively, semi-quantitatively or quantitatively, before the sample enters the flow cell. By means of differential measurements, calibration curves and the like, it is possible in this way to achieve calibration at runtime with just one analysis cell by qualitatively, semi-quantitatively or quantitatively comparing the analyte or analytes to be detected in the sample with the reference substance (s) in the calibration solution can.

However, a preferred embodiment of the cartridge has two identical analysis cells on the chip surface so that analysis of a sample in a first and a known reference substance in the second cell can be performed simultaneously to calibrate / calibrate the analysis method.

For simultaneous and corresponding processing of the sample and the reference substance between the respective outer input and the respective cell, two mutually completely separate channel systems are provided for each other for the same processing in this embodiment. Basically, these channel systems can have different geometrical shapes from each other, because there is only a matter of objectively comparable processing, e.g. in terms of the time course or the amount of added reagents, in the two channel systems. This may be achieved, for example, provided that the sample and reference substances have approximately equal viscosity values by approximately equal lengths of the two channel systems.

In this embodiment, two respective reagent inputs connected to the respective channel system are provided, one of which serves to supply the reference substance and the other to supply air, a buffer solution, enzyme solution, substrate solution or the like as described above for the one-piece system.

Preferably, two or more reagent inputs are provided in the cartridge, by means of which two or more different reagents can be supplied to a channel system. In combination with the above preferred embodiment, the reagent inputs connected to the different channel systems may e.g. be arranged as two parallel rows.

Because of the externally accessible arrangement, two different reagent inputs in the cartridge according to the invention can each be operated independently, e.g. filled with liquid reagents or subjected to negative or positive pressure. Because of their basically similar configuration, however, the different reagent inputs can also be of the same kind, e.g. synchronously, be operated.

As the number of reagent inputs increases, the flexibility, adaptability to the demand of an application, and, if desired, the complexity of the multi-parameter analyzes to be performed with the cartridge, both in terms of reagent selection and analysis process, increases.

In a cartridge according to the invention, a flushing channel may be provided for advancing the sample collected in the sample inlet in the channel system. The flushing channel can open, for example, in the channel system in the sample inlet, possibly with a constriction or a capillary stop (capillary stop) at the discharge point, such that a flushing of the sample can take place only under a pressurization of the flushing channel. In any case, the flushing channel ends on its other side in a reagent inlet for supplying a flushing fluid, for example a buffer solution, or from air.

In specific embodiments with a flushing channel, e.g. Wipe samples are taken (for example in the form of a punched circle) by means of a filter pad placed in the sample inlet and then rinsed or flushed into the channel system. For this, e.g. a syringe module attachment can be used. By means of a syringe module filled, for example, with a rinsing solution (for example a buffer), the sample can be rinsed out of or through the filter pad into the cartridge.

Sampling may be for liquid samples, e.g. for whole blood, by one or more capillaries integrated into the sample inlet. Due to the capillary effect, a capillary is filled with predetermined dimensions, for example, by applying a sample drop, whereby an automatic sample dosing takes place. After closure of the sample inlet with a closure cap, the sample volume thus metered in the cartridge may be e.g. be processed in an automated process.

In the closure cap is housed in a further embodiment, which also comprises a flushing channel, a sealing cap channel which serves in the closed state of the sample substance inlet of a liquid-tight connection between the capillary inlet and the flushing channel. By air or a liquid from the flushing channel, the sample can be displaced from the sample capillary into the channel system for further processing and possibly pre-dilute at the same time. In a simple case, the cap channel has a U-tube shape.

In a channel system according to the invention, a prediluted or else not prediluted sample can subsequently be conveyed from the sample inlet directly into a first dilution and / or waste chamber for optionally further dilution. For this purpose, this chamber can be connected to a reagent inlet for the supply of liquid / air, whereby dilution or rinsing liquid of the chamber e.g. can be supplied via a flushing channel. Alternatively or additionally, the chamber can be connected to a reagent inlet, into which the possibly diluted sample can be sucked in for the purpose of further processing. Further, the chamber may alternatively or additionally be connected to the second opening of a flow cell for receiving the analyzed sample.

Preferably, at least one mixing chamber (in short: mixer) is provided in or in each of the channel system (s) according to the invention, in which the sample or a reference substance can be mixed with one or more reagents on the way to the analysis cell. The respective mixer is for this purpose connected to one or more reagent inputs. In a mixer, for example, a possibly prediluted sample, a buffer, a substrate, an enzyme and / or a detector may be added. These steps can also be divided into different mixers and, for example, be carried out successively.

The measuring chip preferably has an electrical connection point to which an external device for reading the measured data can be connected. The electrical connection (connection) can e.g. be arranged on a side facing away from the surface of the measuring chip provided for the sample analysis side, whereby a liquid contact of the electrical connection can be avoided. This can be realized by electrical vias through the chip to its underside. The electrical structures required for electrical data preparation can be arranged directly in the measuring chip (for example in the form of an application-specific integrated circuit [ASIC]) or implemented in the external device. An ASIC may also be provided on a separate substrate from the measuring chip in the cartridge. For example, the chip may be liquid-tightly mounted on the cartridge by lateral interference fit by means of an elastic, liquid-tight material such as silicone or polydimethylsiloxane (PDMS).

The sampling can be done in a simple case directly by means of the sample input. In the cartridge according to the invention, however, additional means for collecting the sample substance may be accommodated in a separate sampling module. The sampling module includes a sample exit which can be fluid tightly connected to the sample inlet of the cartridge for transferring the sample (e.g., utilizing the capillary forces) into the channel system. In this case, the sampling can be carried out before or after the sampling module is plugged together with the sample inlet. The sampling module is also preferably a low cost disposable article, such as a plastic or glass capillary having a tube end with a rupturable membrane for connection to the sample inlet at one end and a closure cap at the other end. An advantage of a separate sampling module is e.g. in the possibility, for example, in case of a failed sampling only replace this and not the whole measuring cartridge.

In specific embodiments of the cartridge according to the invention, prior to introduction of the sample substance via the sample inlet and / or a reference substance via a reagent inlet, no further substances other than air and / or inert gas, e.g. an inert gas, present in the canal system. Optionally, water, in particular distilled water, may additionally be provided in advance in the channel system of the cartridge.

Another aspect of the invention is a modular disposable measuring cartridge system (short: system) of two separable and zusammenschließbaren main modules: a disposable cartridge according to the invention and a disposable or reusable reagent module with one or more reagent cylinders for receiving reagents, each cylinder via a cylinder outlet for liquid-tight connection to a Reagent input of the cartridge and a displaceable piston (drive piston) for the transport of the sample and the reagents in the system has. The system according to the invention is specified in claim 12.

In the system of the present invention, reagent storage and addition and mechanical means for moving all the gases and liquids in the system (i.e., especially in the cartridge) are housed in a separate module, the reagent module. In general, the use of pasty masses or powdery substances in the reagent module is not excluded.

The system can be connected to an external control and readout device (so-called control unit) with a processor and at least one electro-mechanical actuator for driving the pistons. The control unit is preferably connectable by means of a purely electrical interface with the measuring chip and a purely mechanical interface with the reagent module. In this way, it does not come into contact with the liquids involved in the analysis during normal operation of the system and therefore does not have to be cleaned of these liquids or be serviced solely for this reason.

The reagent module according to the invention is in advance, i. before the actual analysis process, with one or several reagents in liquid form and is prepared e.g. supplied by the manufacturer in a ready-to-use condition or filled on-site by chemically-trained personnel. The term "reagents" in this context includes not only chemically and / or biologically active substances but also water, in particular distilled water, air or other fluids which can be used in an analysis process.

The use of reagent cylinders with pistons in a unitary block allows, on the one hand, a simple, robust and reliable mechanical drive of the entire fluidics, i. all gas and liquid movements in the channel system of the cartridge. On the other hand, it allows an arbitrarily high degree of integration of all fluidic processes in the system, e.g. by a synchronous or suitably coordinated drive of different pistons.

In the assembled state of the system, the pistons can firstly supply reagents to the cartridge and advance them by generating pressure. On the other hand, the pistons may also be pressurized, i. serve as pumps to aspirate or reciprocate substances in the cartridge.

The piston drive according to the invention is in principle similar for all pistons and can take place at a common location, namely a mechanical interface between the reagent module and the control unit. The drive of two or more pistons, for example, by a common electromechanical actuator, such as a motor, or assume several identical actuators in the control unit. For example, the control unit can have an equal number of pressure and / or suction pistons for driving the drive pistons. These can, for example, press the drive pistons directly or pneumatically or, by means of suitable fastening means or in turn, pull them pneumatically against the drive piston, in each case at a cylinder end facing away from the cylinder outlet.

With the basic capability of both independent and properly tuned drive of the various drive pistons in the reagent module, the system of the invention provides virtually unlimited flexibility in multiparameter analysis, both in terms of number and choice of reagents, and timing of an analysis procedure, without any Loss of structural complexity or cost.

In addition, the reagent module according to the invention basically allows visual inspection of its function and integrity, e.g. when using transparent or at least translucent glass or plastic cylinder or such a reagent block. As a result, at least coarse production or process errors, e.g. the clogging of one of the fluidic channels in the system can be detected immediately. This is an advantage over the externally invisible analyzes within miniaturized dry-reagent cartridges hidden in the reader, as known in the art.

With the system according to the invention, the modification or addition of analysis steps, possibly with the addition of other / further reagents, due to the described modular design of the system and the possibility of flexible digital control by a purely electromechanical control unit is extremely easy to implement, because it basically requires none constructive changes in the design of the measuring cartridge. It may already be effected by a change in the control software and possibly obtaining a (one-way) reagent module with other reagents; however, the manufacturing costs or the reproducibility of the analyzes are not impaired.

Furthermore, the principle of construction of the system according to the invention will in principle not become more complex or expensive even with an increasing number of reagent cylinders in the reagent module and / or the channel systems in the cartridge. That the manufacturer can offer several variants of the measuring chip cartridge and reagent modules with different number or arrangement of reagent cylinders or channel systems without switching to other production technologies.

Due to the possibility of complete automation of the entire analysis process from the completed sampling and the same centralized drive In all analysis processes, the system according to the invention ensures maximum reproducibility of the analysis from measurement to measurement.

The invention enables integration and automation of reagent storage, sample collection, sample dilution, reagent processing, detection, and calibration and control functions in a disposable cartridge system for mobile multiparameter analysis.

In a preferred embodiment, the system comprises a cartridge as described above having two identical analysis cells for parallel measurement of a sample and a reference substance, and a reagent module having two reagent cylinder sets corresponding to each other which allow equal processing of the sample and the reference substance. Preferably, these are two identical sets of reagent cylinders, e.g. can be arranged parallel to each other in the reagent block.

In all cases, one or more of the cylinder outlets are sealed in a liquid-tight manner, preferably by a membrane, prior to merging with the cartridge, with the respective reagent inlet (s) of the cartridge each having a hollow needle, e.g. a cannula, for piercing the membrane may have. Preferably, prior to merging with the cartridge, all cylinder exits are each closed with such a membrane, e.g. by a common membrane for a row of cylinders.

The interchangeable modules of the invention can be manufactured inexpensively, e.g. made of plastic by injection molding.

• eine hohe Zuverlässigkeit durch integrierte Kalibrier- und Kontrollfunktionen;
• eine hohe Reproduzierbarkeit der Analysen durch automatisierte Prozessierung in der integrierten Messchipkartusche;
• eine einfache Bedienung (wenig Fehlbedienungsquellen);
• ein robustes System, besonders geeignet für den mobilen Einsatz;
• eine (kosten-)effiziente Verbrauchsmittellogistik durch die einfache Herstellung, Lagerung, Transport der Verbrauchsmittel.

The modular system according to the invention has, inter alia, the following advantages:

• high reliability through integrated calibration and control functions;
• high reproducibility of the analyzes by automated processing in the integrated measuring cartridge;
• a simple operation (few sources of misuse);
• a robust system, particularly suitable for mobile use;
• a (cost) efficient consumption logistics through the simple production, storage, transport of consumables.

1. a) die Probensubstanz am Probeneingang in eine Probenaufnahme, vorzugsweise eine Kapillare, eines unbenutzten Messchipkartuschenmoduls gegeben wird,
2. b) das Messchipkartuschenmodul mit dem Reagenzienmodul flüssigkeitsdicht zusammengeführt wird,
3. c) die Analyse der Probensubstanz durchgeführt wird, wobei die Probensubstanz und die Reagenzien im System mittels der Antriebskolben befördert werden,
4. d) das benutzte Messchipkartuschenmodul vor jeder neuen Probenmessung gegen ein unbenutztes Messchipkartuschenmodul ausgewechselt wird.

The invention is also directed to a method for performing multiparameter analyzes of chemical and / or biological sample substances by means of a system according to the invention, in which

1. a) the sample substance at the sample inlet is placed in a sample receptacle, preferably a capillary, of an unused measuring cartridge module,
2. b) the measuring chip cartridge module is brought together in a liquid-tight manner with the reagent module,
3. c) the analysis of the sample substance is carried out, wherein the sample substance and the reagents are conveyed in the system by means of the drive pistons,
4. d) the used measuring cartridge module is replaced with an unused measuring cartridge module before each new sample measurement.

The above description of the measuring chip cartridge module and of the system according to the invention and the description of the embodiments below with reference to the figures, inter alia, individual features or feature combinations of an operating method according to the invention, if necessary specific to an embodiment, refer.

The system and the method according to the invention can be used everywhere in addition to point-of-care diagnostics (eg for immunoassays) and other medical fields (eg for multi-parameter bioanalyses based on protein tests for mobile use) where a fast, mobile and inexpensive electrochemical analysis of samples is needed.

Preferably, not only the disposable cartridge, but also the reagent module is replaced with an unused one before each new sample measurement. However, this is not mandatory. Rather, a reagent module may be used for two or more sample measurements and may be used between measurements e.g. be separated by the piston liquid-tight from the removable cartridge.

With the system according to the invention, a calibration or control functions can be realized at runtime of the analysis by two analysis cells on a chip (or 2 analysis arrays) for a separate, simultaneous processing of a real sample and a calibration solution (eg a defined analyte solution). be used. Simultaneous and, as far as possible, identical (and as identical as possible) processing of the sample and reference substances may be used, for example, in terms of timing and reagent addition. by synchronously driving the pistons in the sample and reference substance corresponding (e.g., identical) reagent cylinders.

The calibration solution can, for example, in an alternative embodiment of the cartridge with only one analysis cell (see. Figures 5-7 ), also be integrated in a sample buffer. For calibration in such a variant, calibration positions can be provided in the analysis cell, which preferably represents a flow cell. With such a configuration, a calibration can be realized at runtime, as explained above.

A control of the liquid transport via the chip is possible, for example, by measuring the heating power in conjunction with a chip temperature control. For example, changes in the chip temperature (which is regulated to certain setpoints) are monitored by cooling effects when pumping air rather than liquids across the chip. This can a detection of malfunction of the pump (or the piston or its drive) or leaks in the cartridge are used.

The analysis according to the invention can be controlled by an external device, a control unit. Their connections to the system according to the invention are preferably designed such that the control unit does not come into direct contact with the liquids which flow in the system during the analysis. For this purpose, the control unit may e.g. purely electrically connected to the measuring cartridge module and purely mechanically to the reagent module. The analysis process can be completely automated in the composite system controlled by the control unit, wherein the sample substance and the reagents (and / or optionally the reference substance) are transported in the system by means of the driven drive pistons.

• eine fluidische Schnittstelle lediglich zwischen dem Kartuschenmodul und dem Reagenzienmodul (sowie innerhalb des Kartuschenmoduls zum integrierten Chip und ggf. zur Probennahme),
• eine ausschließlich mechanische Schnittstelle zur Steuerungseinheit für den Betrieb der Fluidik im System mittels des Reagenzienmoduls und
• eine ausschließlich elektrische Schnittstelle zur Steuerungseinheit zur Kontaktierung des elektrischen Array-Messchips
• sowie ggf. eine thermische Schnittstelle zu einem zusätzlichen Temperaturmodul, wenn keine "On-Chip-Temperierung" vorgesehen ist,

beim Betrieb des Systems vorliegen.Contamination of a controlling device known to conventional systems by liquids involved in the analysis can be excluded by the system according to the invention by a complete separation of the interfaces from the control unit, by:

• a fluidic interface only between the cartridge module and the reagent module (as well as within the cartridge module for the integrated chip and possibly for sampling),
• an exclusively mechanical interface to the control unit for the operation of the fluidics in the system by means of the reagent module and
• an exclusively electrical interface to the control unit for contacting the electrical array measuring chip
• as well as possibly a thermal interface to an additional temperature module, if no "on-chip temperature control" is provided,

during operation of the system.

To operate a system according to the invention, only a combination of reagent module (storage of liquid reagents) and chip cartridge module (sampling and dilution function plus measurement) is required as the consumable. In contrast to the conventional systems, the possibly existing control unit does not have to be specially serviced, filled, cleaned or replaced.

The removal and disposal of the modules is possible in the form of a closed liquid-tight unit, so that there is no risk of contamination or injury to the user or the patient.

• e lektrische Durchkontaktierungen zur Chipunterseite (optional auch ein 3D-Aufbau: ASIC + Bio-MST) und/oder
• e ine Chipmontage einschließlich fluidischer Abdichtung ggf. durch eine seitliche Presspassung mittels eines elastischen flüssigkeitsdichten Materials (z.B. Silikon, PDMS) möglich.

In chip mounting and contacting, a reduction in the space requirement on the chip (with a concomitant cost reduction) for electrical contacting and fluidic sealing is possible. While in known systems typically both on the Chip top is housed in the system according to the invention in addition to the also possible contacting of the chip top optional

• e lektrische Durchkontaktierungen to the chip bottom side (optionally also a 3D structure: ASIC + Bio-MST) and / or
• a chip assembly including fluidic sealing possibly by a lateral press fit by means of an elastic liquid-tight material (eg silicone, PDMS) possible.

The concrete examples of individual fluidic functions described above and below in a system according to the invention, e.g. The calibration method, sampling, sample dilution, reagent storage and processing should not be construed as exhaustive. The system and operating method according to the invention also allow for combinations of individual fluidic processes and their application-specific variations that are not explicitly described, and which are readily apparent to the person skilled in the art.

Further advantages of the invention are explained below with reference to the embodiments illustrated in the drawings and the associated description. These should have no limiting effect on the subject invention, but serve as specific examples. In particular, the figures are schematic representations that are not to scale.

Fig. 1

ein erstes Ausführungsbeispiel des erfindungsgemäßen Systems mit zwei parallelen Messkanälen für eine Kalibrierung zur Laufzeit der Probenanalyse;

Fig. 2

eine perspektivische Explosionsdarstellung des Beispiels aus Fig. 1 (Ansicht von unten);

Fig. 3

eine perspektivische Explosionsdarstellung des Beispiels aus Fig. 1 (Ansicht von oben);

Fig. 4

Hauptmodule aus Figuren 1-3 im zusammengeführten Zustand;

Fig. 5

ein zweites Ausführungsbeispiel des erfindungsgemäßen Systems mit einem einzigen Messkanal;

Fig. 6

eine perspektivische Darstellung des Beispiels aus Fig. 5 (Ansicht von unten);

Fig. 7

eine perspektivische Explosionsdarstellung des Beispiels aus Fig. 5 (Ansicht von oben);

Fig. 8

eine schematische Darstellung der einzelnen fluidischen Funktionen im System nach dem ersten Ausführungsbeispiel;

Fig. 9

eine schematische Darstellung der einzelnen fluidischen Funktionen im erfindungsgemäßen System nach dem zweiten Ausführungsbeispiel.

Show it:

Fig. 1

a first embodiment of the system according to the invention with two parallel measuring channels for calibration at the time of sample analysis;

Fig. 2

an exploded perspective view of the example Fig. 1 (View from below);

Fig. 3

an exploded perspective view of the example Fig. 1 (View from above);

Fig. 4

Main modules off Figures 1-3 in the merged state;

Fig. 5

a second embodiment of the system according to the invention with a single measuring channel;

Fig. 6

a perspective view of the example Fig. 5 (View from below);

Fig. 7

an exploded perspective view of the example Fig. 5 (View from above);

Fig. 8

a schematic representation of the individual fluidic functions in the system according to the first embodiment;

Fig. 9

a schematic representation of the individual fluidic functions in the system according to the invention according to the second embodiment.

As a measuring chip for the sample analysis can be within the scope of the invention, for example, a biochip of in DE 10 2008 027 038 A1 be used type described. The following description of the specific embodiments shown in the figures therefore relates purely by way of example to such a biochip as a measuring chip in the sense of the invention. Other types of measuring chips, in particular those for purely chemical analyzes, can be used in all cases.

LIST OF REFERENCE NUMBERS

1

piston

2

reagents block

3

reagents cylinder

4

vent hole

5

cartridge

6

connection Adel

7

biochip

8th

cap

9

membrane

10

fluidic

11

Diluent / waste compartment

12

sealing film

13

sample receipt

14a

Flow Cell-A

14b

Flow cell-B

15

piston actuators

16

calibration input

Fig. 1 shows a perspective view of a first embodiment of the system according to the invention with two corresponding measuring channels that allow calibration of the sample analysis at run time. The illustrated integrated biochip cartridge system includes two main interchangeable modules, a reagent storage and processing module (short: reagent module) 1-4, and a biochip cartridge module (generally: cartridge cartridge module, short: cartridge) 5-8.

The individual components are in the exploded views in Fig. 2 (from below) and in Fig. 3 (from above) shown in more detail.

The storage of the various reagents required for the analysis takes place in the reagent cylinders 3 of the reagent block 2. The reagent cylinders are closed by the drive pistons (in short: pistons) 1 and the membranes 9.

The biochip 7 in array format (eg with interdigital electrodes) represents a detection unit for multiparameter analyzes. It is integrated in the measuring cartridge (in short: cartridge) 5 and by means of the flow cells 14a / b with the, or in this example the two separate and corresponding channel systems (hereinafter also: fluidic systems), ie the fluidic channels 10 and the dilution / waste chamber 11 for the sample measuring channel (or the corresponding dilution / waste chamber for the Kalibriermesskanal). A sealing film 12 serves to seal the fluidic channels and the dilution / waste chambers.

For a simple sampling with a defined volume, there are one or more sample capillaries with predetermined dimensions in the sample inlet 13, which is closed after sampling with the closure cap 8. The sample inlet is particularly suitable for receiving drops, e.g. Capillary blood from a fingertip. If the sample substance is in a separate storage container, e.g. a syringe, collected or prepared so it can be supplied by means of a cannula also in the form of drops the sample input. The liquid sample substance then fills the sample capillary (s) due to the capillary effect.

The introduction of the collected sample substance from the sample capillary into the fluidic system of the cartridge takes place, for example, by displacing the sample substance through the air or a rinsing liquid which is supplied to the sample capillary (s) via a channel within the closure cap when the sample inlet is closed, cf. this also the diagram in Fig. 8/9 , Alternatively, the sample substance from the sample capillary can be sucked into the fluidic system of the cartridge by the generation of negative pressure at the capillary outlet by means of a suitable piston in the connected reagent module.

When merging the cartridge module with the reagent module connecting needles or cannulas 6 realize a fluidic connection of the modules by piercing the membranes 9. The processing of the solutions for the realization of Fluidikabläufe during the analysis process via the piston 1, the corresponding reagents from Carry the reagent cylinders 3 into the cartridge. For that, i. during fluidic processing, vent holes 4 are opened. The pistons are thereby in a special device / the control unit (not shown) with the help of automated mechanical drives basically moved independently. In addition to the drive functions for fluidic control, the device also fulfills the read-out functions for the biochip.

The control unit may include, for example, an electromechanical drive for the pistons 1, a processor for the digital evaluation of the measurement signals from the biochip and for controlling the measurement sequences, and a reading device for reading out a measurement protocol software. The control unit, in particular its electrical connections to the cartridge module 5 and mechanical connections to the reagent module, is thereby no direct contact exposed to the processed or processed in the system fluids. This eliminates the maintenance and cleaning measures required for the control unit in the case of known devices, and the cartridge 5 and the reagent block 2 can simply be exchanged after each measurement.

After inserting the reagent block 2 in the cartridge 5, the connection to the control unit and closing the sample input with the cap, the analysis process can be completely automated, controlled by the control unit.

At the end of the analysis process, the cylinder 3 can be closed with the ventilation holes 4 through the corresponding piston to the cartridge 5 out. Thereafter, the cartridge module may be removed and discarded as a closed unit, while the reagent module may be used for further measurement. Alternatively, both exchangeable main modules can be disposed of as a composite and also liquid-tight sealed unit after the measurement ( Fig. 4 ).

Fig. 1 - Fig. 4 show the first embodiment of the invention with a 2-channel calibration function, ie there are two separate fluidic systems, which can be operated synchronously via a coupling in the drive system, for example by a mechanical connection of the piston.

Fig. 8 outlines the corresponding individual, in particular the fluidic functions in a schematic representation. In a fluidic system of the cartridge 5, the sample is first supplied via the sample inlet 13. Controlled by the piston 1, for example by the control unit, thereafter dilution and mixing of the sample with a sample buffer in a first dilution / waste chamber 11. In parallel, in a second fluidic system, a calibration solution with known analyte concentrations in a second dilution / waste chamber rinsed in the perspective view of the Fig. 2 can be seen below the chamber 11.

For processing the sample and the calibration solution through the flow cells 14a / b via the biochip, the corresponding solutions are moved out of the dilution / waste chambers by means of the pistons 1 in the manner of syringe pump drives P1-P6 in the respective fluidic system (or channel system) of the cartridge 5 , The liquids can also be sucked into the cylinders of the reagent module during specific analysis procedures.

Subsequently, a back flushing in the dilution / waste chamber. These processes, as well as the rinsing of the other reagents via the biochip into the waste chamber, take place synchronously for both fluidic systems. The different areas on the biochip, separated by the two separate flow areas, experience an identical process flow. In conjunction with the exact knowledge of the composition and / or Parameters such as concentrations of the reference substance / calibration solution can thus be realized in a precise calibration of the sample analysis at runtime. Furthermore, additional functions, such as positive control positions, in each flow area and control of fluid flow across the biochip, lead to an improvement in quality control in each analysis.

Fig. 5-7 such as Fig. 9 represent a variant of the invention without 2-channel calibration function. The calibration can be realized in such a system via individual calibration positions on the same array in a common flow cell of the measuring chip, as described above.

A significant increase in the degree of integration of the analysis processes in the system according to the invention over the conventional systems by a similar, possibly common, possibly even identical and / or simultaneous control of the piston 1 for reagent and Probensubstanzbeförderung in the fluid channels 10 (the drive takes place possibly simultaneously and identically for the sample and possibly the reference measuring channel) and the sample inlet closed by the closure cap.

The sample volume may e.g. are metered by a precise dimension of the sample capillary which, when immersed in the sample liquid (e.g., in a drop), is completely filled with the sample by capillary forces. When using wipe samples on filter paper of known absorbency, the dosage of the sample substance may e.g. by punching out a predetermined volume of the wiping sample and adding a precisely metered volume of rinsing liquid. In the case of solid powdery sample substances, e.g. Dried blood, a precise dosage can be achieved by filling a container (e.g., integrated with the sample inlet) with a given volume and adding a predetermined volume of solvent or rinse liquid. By a fully automated addition of various reagents from the reagent module into the cartridge, manual dosing by the operator of the system is reduced to nothing or at best to a reproducible minimal action such as filling a visible container with dry powder.

Claims (16)

1. Disposable chip cartridge system for conducting preferably mobile multi-parameter analyses of chemical and/or biological substances, comprising

   a) a replaceable measuring chip cartridge module, comprising

   - a sample inlet for the extraneous introduction of a sample substance into the measuring chip cartridge module,

   - separately thereof at least one reagent inlet for the extraneous introduction of reagents into said measuring chip cartridge module,

   - a measuring chip, the surface of which, as far as being designed for sample analysis, at least partially confines an analysis cell with two openings, such that a liquid comprising said sample substance can pass into the analysis cell through a first opening for the sample analysis, as well as

   - a channel system, in which said sample substance can be conveyed from said sample inlet to said analysis cell and said reagents can be conveyed from the at least one reagent inlet to said analysis cell, wherein said channel system does not comprises any solid reagents, and preferably does not comprise any other substances apart from air and/or inert gas, prior to the introduction of said sample substance via said sample inlet and/or said reagents via at least the one reagent inlet,
   as well as

   b) a replaceable reagent module designed as a uniform block, having two or more reagent cylinders for receiving reagents, wherein each cylinder is provided with a drive piston for conveying said sample substance as well as said reagents in the system and a cylinder outlet for the liquid-tight connection to a reagent inlet of the measuring chip cartridge module.
2. System according to Claim 1, in which said analysis cell of said measuring chip cartridge module is a flow-through cell, such that a liquid comprising said sample substance can pass into said flow-through cell through the first opening and exit said flow-through cell through the second opening.
3. System according to one of the previous claims, in which said measuring chip cartridge module has two identical analysis cells on said surface of said measuring chip designed for the analysis of samples as well as two separate channel systems, corresponding to each other, for simultaneously processing the sample substance and a reference substance for the calibration of the sample analysis, wherein two reagent inlets corresponding to each other are provided, one of which is provided for introducing air or buffers to dilute samples and the other is provided for introducing a reference substance.
4. System according to one of the previous claims, in which said measuring chip cartridge module has two or more reagent inlets for the introduction of two or more different reagents into the channel system.
5. System according to one of the previous claims, in which said channel system of said measuring chip cartridge module has a rinsing channel for rinsing said sample substance from said sample inlet for the further conveying to said analysis cell, wherein one end of said rinsing channel is a reagent inlet for receiving a rinsing fluid, particularly a buffer.
6. System according to one of the previous claims, in which said sample inlet of said measuring chip cartridge module has a capillary for receiving said sample substance as well as an end cap for closing said capillary after taking a sample.
7. System according to Claims 5 and 6, in which said end cap has an end cap channel that, during a closed condition of the sample inlet, is linked to the said capillary in a liquid-tight manner, in order to connect the capillary to said rinsing channel.
8. System according to one of the previous claims, in which said channel system of said measuring chip cartridge module has a first dilution and/or waste chamber, which is connected to the said sample inlet for receiving said sample substance and/or to an inlet for ventilating and/or to a reagent inlet for the further conveying of said sample substance to said analysis cell and/or to said second opening of the flow-through cell of claim 2 for receiving said used sample substance.
9. System according to one of the previous claims, in which said channel system of said measuring chip cartridge module has one or more mixing chambers, in which said sample substance or a reference substance can be mixed with one or more reagents along the way to said analysis cell, and which are connected for this purpose to said sample inlet and/or one or more reagent inlets, respectively.
10. System according to one of the previous claims, comprising a detachable sampling module for collecting said sample substance with a sample outlet, which can be attached to said sample inlet of said measuring chip cartridge module in a liquid-tight manner.
11. System according to one of the previous claims, further comprising

    a) a replaceable measuring chip cartridge module pursuant to claim 3 and

    b) a replaceable reagent module having two, preferably identical sets of reagent cylinders, corresponding to each other.
12. System according to one of the previous Claims, in which one, multiple or all cylinder outlets of said reagent module are connected to a membrane in a liquid-tight manner and the corresponding reagent inlet or inlets of said measuring chip cartridge module has, or have, respectively, a hollow needle for puncturing said membrane.
13. Process for conducting multi-parameter analyses of chemical and/or biological sample substances by means of a system according to one of the claims 1 to 12, in which

    a) said sample substance is fed to said sample inlet,

    b) said measuring chip cartridge module is connected to said reagent module in a liquid-tight manner,

    c) said sample analysis is conducted, wherein said sample substance and/or a reference substance and said reagents are conveyed in the system by means of said drive pistons to said analysis cell(s), and

    d) the used measuring chip cartridge module is replaced with an unused measuring chip cartridge module prior to every new sample measurement.
14. Process according to Claim 13, in which said sample analysis runs automatically controlled by an external device following sampling completed pursuant to step a) in a system composed pursuant to step b), wherein said external device moves said drive piston(s) and/or reads electrical measurement signals of said measuring chip and without coming into contact with fluid in the process.
15. Process according to one of the Claims 13 and 14 using a system according to claim 11 in which a known reference substance is analyzed in the second channel system and the second analysis cell running parallel to that in the first channel system and in the first analysis cell, wherein said sample substance and said reference substance are synchronously subjected to identical processing and measurement by synchronous movement of the respective drive pistons of the two sets of reagent cylinders.
16. Use of the disposable chip cartridge system according to one of the Claims 1 to 12 for point-of-care diagnostics, particularly for immunology tests, and/or for mobile multi-parameter bioanalyses, particularly for blood tests in the case of a mobile application.

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